FLOW CYTOMETRY & HIGH SPEED CELL SORTING: WEGERNER'S GRANULOMATOSIS

该子项目是利用由NIH/NCRR资助的共享仪器补助金提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是资助机构，不一定是研究者的机构。描述（由申请人提供）：UAB流式细胞仪和高速细胞分选设备为UAB的所有nih资助的研究人员提供颗粒和细胞分析的专家协助和仪器。目前的核心设备包括FACS Vantage（已升级为DiVa）， FACS Calibur和FACScan，这些设备是在NIH和该机构的支持下购买的。这些细胞仪在检测荧光染料和荧光颗粒的数量和类型方面能力有限。在本提案中，23名美国国立卫生研究院资助的UAB研究人员申请资金购买BD LSR II型细胞仪。LSR II将配备一个固态488nm蓝色激光器（20mw）、JDS单相633nm红色激光器（18mw）、405nm紫色激光器（25mw）和一个532nm绿色激光器（可变0-150 mw）。该系统将能够探测到来自蓝色激光的两种颜色加上三角光学阵列的侧面和正向散射，来自红色激光的三种颜色利用三角光学阵列，来自紫色激光的八种颜色利用八角形光学阵列，来自绿色激光的五种颜色利用八角形光学阵列。该系统能够采集多达20个参数。收集到的发射光将通过光纤技术传送到光学阵列。适当的带通滤波器和二向色镜包括在内。LSR II细胞仪将提供目前UAB研究人员无法获得的三种技术：广泛光谱荧光染料的多参数分析；多报告结构的激发和发射检测；量子点荧光发射的激发与检测。该提案记录了共同研究人员使用扩展技术能力的实验计划，缺乏可获得的替代品，以及对该细胞仪的机构管理和物质支持。结合该机构在研究者教育和推广方面的努力，该提案将显著改善流式细胞仪服务，以支持NIH在UAB资助的项目。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): The UAB Flow Cytometry and High-Speed Cell Sorting Facility provides expert assistance and instruments for analysis of particles and cells that is available to all NIH-funded researchers at UAB. The current equipment in the core includes a FACS Vantage (since upgraded to DiVa), a FACS Calibur and a FACScan, which were purchased with support from both the NIH and the institution. These cytometers have limited capabilities in the number and type of fluorochromes and fluorescent particles they can detect. In this proposal, twenty-three NIH-funded, UAB investigators request funds for purchase of a BD LSR II cytometer. The LSR II will be equipped with a solid-state 488nm blue laser (20mw) a JDS Uniphase 633nm Red laser (18mw) and 405nm Violet laser (25mw) and a 532nm Green laser (Variable 0-150 mw). The system will have the ability to detect 2 colors from the blue laser plus side and forward scatter utilizing a trigon optical array, 3 colors from the red laser utilizing a trigon optical array, 8 colors off the violet laser utilizing an octagon optical array and 5 colors off the green laser utilizing an octagon optical array. The system is capable of collecting up to 20 parameters. The collected emission light will be delivered to the optical arrays by fiber optic technology. Appropriate bandpass filters and dichroic mirrors are included. The LSR II cytometer will provide access to three technologies not currently available to UAB researchers: Multi-parameter analysis of a wide spectrum of fluorochromes; Excitation and detection of emission for multiple reporter constructs; Excitation and detection of fluorescent emission from quantum dots. This proposal documents the experimental plans of the co-investigators for use of the expanded technical capabilities, the absence of accessible alternatives, and the institutional administrative and material support for this cytometer. Combined with efforts by the Facility in investigator education and outreach, this proposal will significantly improve the flow cytometry services in support of NIH funded projects at UAB.